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है”ह”ह

IS 3752 (2005): Alcoholic drinks - Methods of test [FAD 14:Drinks and Carbonated Beverages]

IS 3752:2005

Indian Standard

ALCOHOLIC DRINKS — METHODS OF TEST

(Second Revision)

ICS 67.160.10

.,. ..‘4

@ BIS 2005

BUREAU OF INDIAN STANDARDSMANAK BHAVAN, 9 BAHADUR SHAH ZAFAR MARG

NEW DELHI 110002

Jufy 2005 Price Group 7

Drinks and Carbonated Beverages Sectional Committee, FAD 14

FOREWORD

This Indian Standard (Second Revision) was adopted by the Bureau of Indian Standards, after the draft finalizedby the Drinks and Carbonated Beverages Sectional Committee had been approved by the Food and AgricultureDivision Council.

This standard was originally issued in 1967 and revised in 1988.

This revision has been taken up in order to update the methods of test in the light of experience gained in itsusage and align it with the latest practices being followed in the field in the country.

In the preparation of this standard, due consideration has been given to the : (a) Prevention of Food AdulterationAct, 1954 and the Rules framed thereunder (b) Standards of Weights and Measures (Packaged Commodities)Rules, 1977; and (c) State Excise Duty Ru[es which permit the withdrawal of duty free samples for testing. It isrecommended that sample for testing by Bureau of Indian Standards, whenever called for, may also be exemptedfrom excise duty. The standard is subject to restrictions imposed under these Rules, wherever applicable.

For the purpose of deciding whether a particular requirement of this standard is complied with, the finalvalue, observed or calculated, expressing the result of a test or analysis, shall be rounded off in accordance withIS 2:1960 ‘Rules for rounding off numerical values (revised)’. The number of significant places retained in therounded off value should be the same as that of the specified value in this standard.

i

I

IS 3752:2005

Indian Standard

ALCOHOLIC DRINKS — METHODS OF TEST

(Second Revision)

1 SCOPE

This standard prescribes the methods of test foralcoholic drinks. For the estimation of esters, higheralcohols, aldehydes, furfural and methanol, gaschromatographic (GC) method is provided in AnnexA as an alternative method.

2 REFERENCES

The standards listed in Annex B contain provisions,which through reference in this text, constituteprovision of this standard. At the time of publication,the editions indicated were valid. All standards aresubject to revision, and parties to agreements based onthis standard are encouraged to investigate thepossibility of applying the most recent editions of thestandards indicated in Annex B.

3 QUALITY OF REAGENTS

Unless specified otherwise, pure chemicals shall beemployed in tests and distilled water (see IS 1070)shall be used where use of water as reagent isintended.

4 DETERMINATION OF ETHYL ALCOHOLCONTENT

Three methods have been prescribed, namely, Method1, Method 2 and Method 3. Method 1 shall be used asroutine method for apparent strength, Method 2 asreferee method for real strength, and Method 3 in caseof those alcoholic drinks which yield volatile oils ondistillation.

4.1 Method 1, Hydrometer Method

4.1.1 See 1S 2302 and express the strength as percentby volume at 20120”C.

4.2 Method 2, Pyknometer Method

4.2.1

a)

b)

c)

d)

Apparatus

Distillation assembly, the delivery end of thecondenser is attached to a glass tube with a bulbby means of a ground glass joint. The lowerpart of this tube should reach the bottom of thereceiver and dip into the minimum quantity ofdistilled water.

Pyknometer, 25 to 50-ml capacity.

Thermometer, O to 50°C.

Measuring @ask, 200-ml capacity.

4.2.2 Procedure

4.2.2.1 Take 200 ml of sample in a 500-ml distillationflask containing about 25 ml of distilled water and afew pieces of pumice stone. Complete the distillationin about 35 min and collect the distillate in a 200 mlmeasuring flask till the volume in the flask nears themark. Allow the distillate to come to roomtemperature, make up the volume to 200 ml withdistilled water and mix thoroughly.

4.2.2.2 Find out the specific gravity of the distillateat the required temperature with the help of aPyknometer. Obtain corresponding alcohol, percent byvolume, from the tables given in IS 3506.

4.3 Method 3, Distillation Method

4.3.1 Distillation Assembly

4.3.2 Apparatus

a) A4easuringj7ask, 200-ml capacity.

b) Separating jiwnels, 500-ml capacity.

4.3.3 Reagents

4.3.3.1 Sodium chloride, powder.

4.3.3.2 Petroleum ether, 40 to 60”C.

4.3.3.3 Sodium hydroxide, 0.1 N.

4.3.3.4 Phenolphthalein, powder.

4.3.4 Procedure

4.3.4.1 Measure 200 ml of the liquor sample in ameasuring flask. Transfer to a separating funnel,wash, the measuring flask with about 100 ml of water,add the washings to the content of the separatingfunnel and add sufficient powdered sodium chlorideto saturate the liquid. Add about 100 ml of petroleumether and shake vigorously for 2 to 3 min. Allowthe mixture to stand for 15 to 30 min and run thelower layer into a distillation flask. Wash the petroleumlayer twice with about 20 ml of the saturatedsolution of sodium chloride. Add these washings tothe distillation flask. Make the mixed solutionsjust alkaline with sodium hydroxide solutionusing phenolphthalein powder as indicator, add alittle pumice powder and distil. Collect the distillatein the measuring flask till it nears the mark. Bringthe distillate to room temperature, make up the

.,,.

.

i

1

.,.,

1S 3752:2005

volume to 200 ml with distilled water and mixthoroughly.

6.3 Calculation

Calculate ash as follows:100 (ln2- ??$)

Ashcontent, percent (ndv) =v

4.3.4.2 Find outthespecific gravity of the distillateat a particular temperature with the help of aPyknometer. Obtain corresponding alcohol, percent byvolume from the tables given in IS 3506.

where

rnz= weight of the dish with ash, in g;5 DETERMINATION OF RESIDUE ONEVAPORATION

m, = weight of the empty dish, in g; and

v = volume of liquor taken for estimation, in ml.5.1 Apparatus

7 DETERMINATION OF TOTAL ACIDITY5.1.1 Hot Air Oven

Two methods have been prescribed, namely,Method 1 and Method 2. Method 1 shall be used forthe products which are colorless like Vodka and Gin.Method 2 shall be used for the products which arecoloured.

5.1.2 Water Bath

5.1.3 Measuring Flask, 200-ml capacity.

S.1.4 Dish, 150-ml capacity, silica or platinum.

7.1 Method 15.1.5 Desiccator

7.1.1 Reagents5.2 Procedure

7.1.1.1 Standard sodium hydroxide, 0.05 N.Evaporate 200 ml of the sample in a dried, tared dishon a water bath. Dry the dish in an air oven at 110 +2°C. Cool in desiccator and weigh the dish. Repeat tillconstant weight is obtained. Calculate the dissolvedsolids to fourth decimal places.

7.1.1.2 Phenolphthalein indicator, as prescribed inIS 2263.

7.1.2 Procedure

Take 50 ml of sample and add about 200 ml ofneutralized distilled water. Titrate against standardsodium hydroxide solution using phenolphthalein asindicator.

5.3 Calculation

Calculate residue on evaporation as follows:

100 (m, – m,)Residue on evaporation, =

percent (m/v) v 7.1.3 Calculation

Calculate the total acidity as follows:where

m2= weight of the dish with dry material, in g;

m, = weight of the empty dish, in g; and

v = volume of liquor taken for estimation, in ml.

Total acidity expressed as tartaric acid, grams per100 litres of absolute alcohol =

~xooo0375x looxlof)ox2

v, .6 DETERMINATION OF ASH

where

V = volume of standard sodium hydroxide used fortitration, in ml; and

V, = alcohol, percent by volume.

6. I Apparatus

6.1. I M@e Furnace

6.1.2 Desiccator7.2 Method 2

6.1.3 Water Bath7.2.1 Apparatus

7.2.1.1 pH meter6.1.4 Dish, 150-ml capacity, silica or platinum.

6.2 Procedure7.2.1.2 Magnetic stirrer

7.2.2 Reagents

7.2.2.1 Standard sodium hydroxide, 0.05 N.

7.2.2.2 Bu~er solutions, ofpH = 4.0, 7.0 and 9.2.

7.2.3 Procedure

Shake the content of the container. Evaporate 100 mlof the sample in a dried, tared dish on a water bath.Place the dish with the residue on a hot plate or on alow flame; heat the contents at low temperature untilthoroughly charred. Place the dish in the muffle furnacemaintained at 450° to 500°C for about an hour. Coolthe dish in a desiccator and weigh. Express the resultto fourth decimal places. Calibrate and standardize thepH meter using the buffer

L

!,

,’.

solutions ofpH = 4.0, 7.0 and 9.2. Add approximately100 ml of distilled water to 250-ml beaker. Place onestirrer in the beaker and place it on the magnetic stirrer.Carefully immerse the electrode into the beaker. Addapproximately 50 m}of sample and rapidly titrate withstandard sodium hydroxide solution to pH 8.2. Add50 ml of sample into the resulting solution and titrateto pH 8.2. Record the volume of standard sodiumhydroxide solution consumed for the secondtitration.

7.2.4 Calculation

Calculate the total acidity as follows:

Total acidity expressed as tartaric acid, grams per 100litres of absolute alcohol=

~xO.003Tsx100xlooox2

v,where

V = volume of standard sodium hydroxide used fortitration, in ml; and

VI = alcohol, percent by volume.

NOTE — One millilitre of standard sodium hydroxide is

equivalent to 0.00375 g oftartaric acid.

8 DETERMINATION OF VOLATILE ACIDITY

8.1 Reagents

8.1.1 Standard Sodium Hydroxide, 0.05 N.

8.1.2 Phenolphthalein Indicator, as prescribed in1S2263.

8.2 Procedure

Take 50 ml of the distillate obtained from thedetermination of ethyl alcohol (see 4.2.2.1) and titrateagainst standard sodium -hydroxide solution usingphenolphthalein as indicator.

8.3 Calculation

Calculate the volatile acidity as follows:

Volatile acidity expressed as acetic acid, grams per100 Iitres of absolute akohol =

VXO.OO3X1OOX1OOOX2

v,

where

V = volume of standard sodium h~roxide used fortitration, in ml; and

V, = alcohol, percent by volume.

NOTE — One millilitre of standard sodium hydroxide is

equivalent to 0.003 g of acetic acid.

9 DETERMINATION OF FIXED ACIDITY

9.1 Reagents

9.1.1 Standard Sodium Hydroxide, 0.05 N.

9.1.2 Phenolphthalein Indicator, as prescribed in1S2263.

9.2 Procedure

Take 50 ml of sample and evaporate to near dryness.Dilute with neutralized distilled water to 100 ml andagain evaporate to dryness. Dilute and titrate againststandard sodium hydroxide solution usingphenolphthalein as indicator.

9.3 Calculation

Calculate fixed acidity as follows:

Fixed acidity expressed as tartaric acid, grams per 100Iitres of absolute alcohol =

VX().()()3T5X1()OX1()()OX2

v,where

V = volume of standard sodium hydroxide used fortitration, in ml; and

VI = alcohol, percent by volume.

NOTE — One millilitre of standard sodium hydroxide is

equivalent to 0.00375 g of tartaric acid.

10 DETERMINATION OF ESTERS

10.1 Reagents !., ..*,

10.1.1 Standard Sodium Hydroxide, 0.1 N.‘*

10.1.2 Standard Sulphuric Acid, 0.1 N.

10.2 ProcedureI

10.2.1 To the neutralized distillate from the volatileacidity determination (see 7.2), add 10 ml of standardalkali and reflux it on a steam bath for an hour. Cooland back titrate the excess of alkali with standardsulphuric acid.

10.2.2 Simultaneously run a blank taking 50 ml ofdistilled water in place of the distillate of the sample tin the same way. The difference in titration value inmillilitres of standard acid solution gives the equivalentester.

10.3 Calculationt

Calculate the esters as follows:

Esters expressed as ethyl acetate, grams/100 Iitres ofabsolute alcohol =

Vxo.fJ088x looxlofjox2

v,

where

V= difference of standard acid used for blank andsample, in ml; and

VI = alcohol, percent by volume.

NOTE — One millilitre of standard alkali is equivalent to

0.0088 g of ethyl acetate.

3

1S 3752:2005

11 DETERMINATION OF HIGHER ALCOHOLS

Three methods have been prescribed, namely, Method1, Method 2 and Method 3. Method 1 maybe used todetermine approximately the quantity of higheralcohols in process control. Method 2 shall be employedfor accurate determination, while Method 3 shall beused as referee method.

11.1 Method 1 (Komarowski Method)

11.1.1 Reagents

a) Salicylic ukiehyde, 1 percent (tn/v) — Dissolve1g salicylic aldehyde in 95 percent alcohol (freeof higher alcohol) to make 100 ml.

b) Sulphuric acid, conforming to IS 266.

11.1.2 Procedure

11.1.2.1 Take a clean glass-stoppered bottle and washit twice with the sample to be tested. Similarly, wash asmall graduated cylinder or a 100-ml pipette.

11.1.2.2 Take 10 ml of sample in the bottle and add1 ml of 1 percent salicylic aldehyde and 20 ml ofconcentrated sulphuric acid. Allow to stand at roomtemperature for over 12 h. Note the colour change(For quick routine analysis, the colour change maybenoted after a shorter interval of about 15 to 20 min at15° to 20”C).

The colours developed after the reaction indicate theamount of higher alcohol as follows:

Colour Amount of Higher Alcohol

Light yellow Only traces

Yellow to brownish About 0.01 percent (v/v)

Brown 0.02 to 0.03 percent (v/v)

Red About 0.05 to 0.1 percent(v/v)

Dark red to black Above 0.1 percent (v/v)

11.2 Method 2

11.2.1 Apparatus

It consists of an 800-ml capacity Kjeldahl flaskfitted with splash head connected to distillation head.The other end of distillation head is connected to aLiebeg condenser, which opens into a receiver havinga capacity of 250 ml. All joints are made of groundglass.

11.2.2 Reagents

a) Sulphuric acid, conforming to IS 266.

b) Oxidizing mixture, Dissolve 100 g of potassiumbichromate in 500-ml distilled water and add100 ml of sulphuric acid and make the volumeto 1 Iitre with distilled water.

c) Standard sodium hydroxide, 0.1 N.

d)

e)

f)

g)

Carbon tetrachloride, distilled.

Sodium chloride, AR grade.

Sodium sulphate, AR grade.

Phenolphthalein indicator, asIS 2263.

11.2.3 Procedure

11.2.3.1 Add 50 ml of water to

prescribed in

the solutionresulting from the determination of esters (see 10.2.1).Saturate it with sodium chloride; extract the saturatedsolution four times using 40, 30, 20 and 10 ml,respectively of carbon tetrachloride. Combine theextracts and wash three times with saturated sodiumchloride solution and twice with saturated sodiumsulphate solution.

11.2.3.2 Filter the extract and add 50 ml of oxidizingmixture. Reflux for 2 h. Cool, wash the refluxcondenser with 50 ml of water and transfer it todistillation flask using about 50 ml of ivater. Distil tillabout 50 ml is lefl over in the flask. Avoid charring.

11.2.3.3 Titrate the distillate against standard sodiumhydroxide, using phenolphthalein as indicator.

11.2.3.4 Run a blank in the same way taking 50 mlof distilled water in place of the distillate of theliquor.

11.2.4 Calculation

Calculate higher alcohol as follows:

Higher alcohol expressed as amyl alcohol, grams per100 litres of absolute alcohol =

Vxo.()()ggxlf)()xlo()()xz

q x V2

where

V= difference of standard acid used for blank andsample, in ml;

V{ = volume of sample taken; and

Vz = alcohol, percent by volume.

NOTE — One millilitre of standardalkali is equivalent to

0.0088 g of amyl alcohol.

11.3 Method 3

11.3.1 Apparatu~ Spectrophotometer.

11.3.2 Reagents

a)

b)

c)

p-dim ethylaminobenzaldehyde solution —Dissolve 1 g p-dimethylaminobenzaldehyde inmixture of 5 ml sulphuric acid and 90 ml waterin a 100-ml volumetric flask and dilute to markwith water.

Isobutyl alcohol, AR grade.

Isoamyl alcohol, AR garde.

‘1

.

4

d)

e)

Ethyl alcohol, redistilled, middle 50 percentfraction.

Synthetic standard higher alcohol, weigh 2 gisobutyl alcohol and 8-g isoamyl alcoho[into iIitre volumetric flask and dilute to mark withwater. Pipette two 10 ml portions into 100-mlvolumetric flasks and dilute to mark, one withwater and other with ethanol. Prepare workingstandards for products in the range of Oto 170proof containing 1.0 to 6.0 g synthetic higheralcohol per 100 Iitres by diluting 1.0 to 6.0 mlaliquots of aqueous standard solution to 100 mlwith alcoholic solution of proof expected fordiluted sample when pipetted into analysis tube.Prepare similar working standards for productsin range of 170 to 190 proof by diluting 1.0 to6.0 ml aliquots of alcohol standards solution to100 ml with alcohol solution of number ofsamples or its dilution. When 6 ml syntheticstandard diluted with 190 proof alcohol iscarried through analysis, absorbance shall be0.83 * 0.03 at 530 nm.

11.3.3 Procedure

11.3.3.1 Preparation of sample

a)

b)

Take 200 ml of sample (liquor) in 500-mlErlenmeyer flask, add about 35 ml water andfew grains of Carborundum. Distil slowly into200-ml volumetric flask until distillate is nearlyat mark. Make up to mark with distilled waterand mix.

For samples containing 6 g fusel oil per 100litres, dilute the distill~d sa~ple with water toconcentration of 2.0 to 5.0 g fusel oil per 100litres (dilute 5 ml brandy, rum, or blendedwhisky to 100 ml; dilute 5 ml blended brandy,rum or straight whisky to 250 ml).

Pipette 2 ml of aliquot of sample or diluted sample,2. ml of water (for reagent blank), and 2 ml aiiquotof standards into 15 mm x 150 mm stoppered orcovered test tubes. Stopper or cover tubes, placein rack, and then in ice-bath. Pipette 1 mlp-dimethylaminobenzaldehyde solution into eachtube; shake, and replace in ice-bath for 3 min. Withtubes still in ice-bath, add 10 ml sulphuric acid fromburette down the side of the each tube. Shake tubesindividually and replace in ice-bath for 3 min. Transferrack of tubes from ice-bath to boiling water-bath for3 to 5 min and then to room-temperature-bath. Readpercentage transmittance of developed colour ofsamples and standards on spectrophoton-teter at 530to 535 pm against reagent blank as reference (use samewavelength for both standards and unknowns). Plot ghigher alcohol per 100 litres on linear scale of asabscissa against percentage T as ordinate on log scaleof Semi log paper. Convert percentage transmittanceof samples to g higher alcohol per 100 Iitres from

IS 3752:2005

standard curve. If dilution was used, multiply g higheralcohol per 100 litres in original sample. Analyze twolevels of standards with each series of unknowns.

12 DETERMINATION OF ALDEHYDES

12.1 Apparatus

12.1.1 Iodine Flask, of 250-ml capacity.

12.2 Reagents

12.2.1 Sodium Bisulphite Solution, approximately0.05 N.

12.2.2 Standard Iodine Solution, 0.05 N.

12.2.3 Standard Sodium Thiosulphate Solution,0.05 N.

12.2.4 Starch Indicator, as prescribed in IS 2263.

12.3 Procedure

Take 50 ml of distillate of liquor (see 4.2.2.1) in a250-ml iodine flask and add 10 ml of bisulphitesolution. Keep the flask in a dark place for 30 minwith occasional shaking. Add 25 ml of standard iodinesolution and back titrate excess iodine against standardsodium thiosulphate solution using starch indicator.Run a blank taking 50 ml of distilled water in place ofdistillate of the liquor in the same way. The differencein titration value, in millilitres, of sodium thiosulphatesolution gives the equivalent aldehyde.

12.4 Calculation

Calculate aldehydes as follows:

Aldehydes expressed as acetaldehyde, grams per 100litres of absolute alcohol =

J7xo.()()ll xl(j()xl()()()xz

VI

where

V = difference of standard sodium thiosulphatesolution used for blank and sample, in ml; and

VI = alcohol, percent by volume.

NOTE — One millilitre of standard sodium thiosulphate

solution is equivalent to 0.001 1 g of acetaldehyde.

13 DETERMINATION OF FURFURAL

13.1 Apparatus

Nessler tubes with flat bottom tubes of thin colorlessglass 25 mm in diameter, about 150 mm in length andgraduated at 50 ml. The depth measured internally fi-omgraduation mark to the bottom shall not vary by morethan 2 mm.

13.2 Reagents

13.2.1 Aniline, distilled and colorless.

5

IS 3752:2005

13.2.2 Hydrochloric Acid, conforming to IS 265.

13.2.3 Furfural Free Alcohol — Let alcohol containing5 g ofrn-phenylene diamine hydrochloride per litrestand at least for 24 h with frequent shaking (previoustreatment with potassium hydroxide is not necessary).Reflux for at least 8 h, longer if necessary. Let standovernight and distil, rejecting the first 100 ml and thelast 200 ml of the distillate. If this gives colorationwith aniline hydrochloride, repeat the treatment.

13.2.4 Standard Furfural Solution — Dissolve 1 g ofredistilled, colorless furfural in 100 ml of furfural freealcohol (see 13.2.3). Prepare standard furfhral solutionby diluting 1 ml of this solution to 100 ml with 50percent furfural free alcohol. One millilitre of thisdiluted solution contains 0.1 mg of furfural (strongfurfural solution will retain its strength but the dilutedstandard solutions should be prepared afresh everytime).

13.3 Procedure

13.3.1 Take 5 ml of distillate (see 4.2.2.1), add 1 mlof colorless aniline and 0.5 ml of hydrochloric acid,and keep for 15 min. Red colour indicates the presenceof furfural. Proceed for quantitative estimation asin 13.3.2, if colour develops.

13.3.2 Dilute a measured portion of the distillate with50 percent furfural-free alcohol to 50 ml. First add 2mIof colorless aniline and then 0.5 ml of hydrochloricacid. Mix and keep at 15°C for 15 min. Compare thecolour developed with standard furfural solution.

13.4 Calculation

Calculate furfural as follows:

Furfural, grams per 100 Iitres ofabsolute alcohol =

W)xloooxlo oxloo

l“,x V2where

w = weight of furfural present in volume used formatching the experimental solution, in g;

VI = volume of experimental solution used forestimation, in ml; and

V2= alcohol, percent by volume.

14 DETERMINATION OF COLIFORM COUNT

Two methods are prescribed, namely, Method 1 andMethod 2. Method 1 may be used as routine methodand Method 2 as referee method.

14.1 Method 1

Coliform bacteria include all aerobic and facultativeanaerobic Gram-negative non-spore-forming bacteria,which ferment lactose with the production of acid and

gas. A positive presumptive test is indicated by theformation of acid and gas within 48 h at 35 to 37°C ina fermentation tube containing lactose bile salt broth.Alternatively, the development of dark red coloniesapproximately 0.5 mm in diameter in a solid medium(violet red bile agar) within 20 to 24 h at 35 to 37°Cmay be considered as a positive evidence for thepresence of coliforrn bacteria. Violet red bile agar isone of the standard media used for the determinationof general types of coliform organisms, including thoseof faecal origin in water, milk and other materials ofsanitary importance.

14.1.1 Apparatus

a)

b)

c)

Weighing scoop, sterile, with counter-weight.

Bacteriological pipettes, sterile accuratelygraduated with cotton plug i’n the upperorifice.

Dilution bottles, sterile, made of heat-resistantglass (preferably borosilicate glass), closed withrubber stoppers (preferably screw caps) with newfriction-fit liners for making them leak-proofand of the following capacities:

1) 150 ml, with mark at 99 ml level; and

2) 25 ml, with mark at 9 ml level.

d) Petri dishes, with outside dish diameter 100 mm,inside dish diameter 91 mm and depth 15 mm.The exterior and interior surface of the bottomshould be flat and free from bubbles, scratchesor other defects, which would interfere with thecounting of the colonies.

e) Bacteriological tubes, sterile, 25-ml capacitywith a mark at 10-ml level and with cottonplugs.

~ Durham fermentation tubes.

14.1.2 Reagents

a) Dissolve 34 g of potassiumphosphate in 500 ml bf distilled

dihydrogenwater, adjust

the pH to 7.4 with 1 N sodium hydroxidesolution and make up to 1 litre with distilledwater. Dilute 1.25 ml of this stock phosphatebuffer solution with distilled water to 1 Iitre toobtain dilution water.

b) Medium — Violet red bile agar shall be themedium, the composition of which shall be asfollows, its final pH being 7.4+ 0.1:

Yeast extract 3.0 g Lactose 10.0 g

Peptone 7.0 g Agar-agar 20.0 g

Sodium 1.5g Sodium chloride 5.3 gtaurocholate

Water 1000 ml

6

1..

Indicator :

1) Neutral red 0.03 g

2) Crystal violet 0.002 g

14.1.3 Procedure

a)

b)

Dilution — Measure 1 ml of the materialfrom the individual samples using a sterilepipette and transfer into 99 ml of dilute water.Mix the contents properly. Prepare serialdilutions of this and add 1 ml of suitabledilutions in triplicate to the sterile petridishes.

Pouring plates — Melt the medium [see14.1.2 (b)] in bacteriological tubes and keep at48° to 50°C. Introduce this medium asepticallyat 42° to 44°C into the petri dishes and mix byrotating and tilting dishes without spreadingover the edges. Spread the mixture evenly overthe bottom of the plate. Allow,to solidify. Atlersolidification of medium in plate, add cover layerof medium.

c) Incubation — Invert the plates and incubate at35° to 37*C for 24 h.

d) Counting — Count the dark red colonies withdiameter of approximately 0.5 mm.

e) Computation — Compute the coliformcount per millilitre from the dilutions used[see 14.1.3 (a)].

NOTES

1 In case of doubt regarding the colonies developed on violet

red bile agar, representative colonies are picked and transferred

to lactose bile salt broth in Durham fermentation tubes.

Production of acid and gas is confirmatory for coliform

organisms.

2 All precautions shall be observed to prevent microbiological

contamination throughout the test.

14.2 Method 2

This method consists of inoculating the beverage intoMacConkey broth-medium. A positive presumptive testis indicated by the formation of gas within 48 h at37°C in the fermentation tube containing the abovemedium. Subsequently, the production of gas inbrilliant green lactose bile broth may be considered asa positive evidence for the presence of coliformbacteria.

14.2.1 Apparatus, see 14.1.1.

14.2.2 Reagents

a) Water, distilled.

b) MacConkq broth: Composition of MacConkeybroth medium shall be as follows, its final pHbeing 7.2 * 0.1:

IS 3752:2005

Peptone 20.0 g Sodium 0.5 gtaurocholate

Lactose 10.0 g Sodium chloride 5.0 g

Water 1000 ml

Dissolve the above ingredients in distilled water.Add 5 ml of 0.04 percent bromocresol purpleper 95 ml of the medium. Fill 10 ml in each ofthe bacteriological tubes. Sterilize in anautoclave for 15 min at 1 kgf/cm2 pressure(121”C).

c) Brilliant green lactose bile broth: Compositionof brilliant green lactose bile broth medium shallbe as follows, its final pH being 7.2* 0.1.

Peptone 10.0 g Bacto oxgall 20.0 g

Lactose 10.0 g BrilIiant green 0.0133g

Water 1000 ml

Dissolve the above ingredients in distilled water.Sterilize in an autoclave for 15 min at 1 kgf/cm2pressure (121”C).

14.2.3 Procedure

Inoculate the beverage into the tubes containingMacConkey broth at 35° to 37”C. If gas is producedwithin 48 h, transfer the broth to the tubes containingbrilliant green lactose bile broth [see 14.2.2(c)] forfurther confirmation of the presence of coliformbacteria. The latter medium inhibits the growth of thelactose fermenting bacteria other than coliforms andhence the production of gas in it positively shows thepresence of coliform bacteria.

15 DETERMINATION OF COPPER

Two methods, namely, diethyldithiocarbamate methodand potassium ferrocyanide method are employed. Thepotassium ferrocyanide method is easier to perform andsufficiently sensitive and accurate for routine type ofanalysis. The diethyldithiocarbamate method (see 15.3)is more sensitive and shall serve as a referee methodin case of dispute or whwe zinc is present.

15.1 Apparatus

15.1.1 Nessler Tubes — Flat bottom tubes of thin,colorless glass, about 25 mm in diameter and about150 mm in length and graduated at 50 ml: The depthmeasured internally from graduation mark to thebottom shall not vary by more than 2 mm in the tubesused for the test.

15.2 Reagents

15.2.1 Dilute Sulphuric Acid, approximately 10percent (vIv).

15.2.2 Aqua Regia, a mixture of one volume of

7

IS 3752:2005

concentrated nitric acid, and three volumes ofconcentrated hydrochloric acid. Nitric acid shall beconform ing to IS 264 and hydrochloric acid shall beconforming to 1S265.

15.2.3 Citric Acid, AR grade.

15.2.4 Dilute Ammonium Hydroxide, approximately10 percent (v/v).

15.2.5 Standard Copper Solution — Dissolve 1.119 gof copper sulphate (CuSOd.5H20) in water and diluteto one litre. Dilute 10 ml of this solution to 100 ml.One millilitre of the diluted solution contains0.02845 mg of copper. The diluted solution shallalways be prepared immediately before use.

15.2.6 Sodium Diethyldithiocarbamate — PrepareO. 1 percent by weight solution of sodiumdiethyldithiocarbamate in water. Sometimesdiethyldithiocarbamate available may not becompletely soluble in water, in which case the insolublematerial may be removed by filtration through anash less filter paper. The reagent is best preparedjust before use, but may stand for one or two weeksin amber coloured bottle without appreciabledeterioration.

15.2.7 Carbon Tetrachloride, AR grade.

15.2.8 Ammonium Chloride, AR grade.

i 5.2.9 Acetic Acid, approximately 5 percent by weight.

15.2.10 Potassium Ferrocyanide Solution,approximately 4 percent by weight.

15.3 Diethyldithiocarbamate Method

15.3.1 In the presence of copper, an aqueous solutionof sodium (or zinc) diethyldithiocarbamate gives agolden brown colour in acid or ammoniacal or neutralsolution. The diethyldithiocarbamate method hasadvantages over the ferrocyanide method, which is invogue in some laboratories since it is more sensitiveand is free from interference by iron and zinc. Thismethod is suitable when the copper content ranges from0.01 to 0.15 mg of copper in the quantity of the materialtaken. With larger quantities of copper, the mixture ofthe test solution and reagent rapidly becomes cloudyand any observance of this in the prescribed test issufficient for condemning the sample as containingexcessive quantities of copper. If a quantitativedetermination is required, the test should be repeatedby using proportionately smaller quantities of samplefor test.

15.3.2 There are two variations of the method, thatis: (a) without extraction, and (b) with extraction.Ordinarily, it is not necessary to resort to the procedureof extraction, which shall be obligatory only when a

15.3.3 Preparation of Test Solution — Transfer20 ml of the material into silica evaporating dish andadd 1 ml of dilute sulphuric acid. Heat gently in thebeginning and then evaporate almost to dryness on awater-bath. Ignite the residue over a smokeless flameto eliminate sulphuric acid. Cool, dissolve the residuein 2 ml of water, add three drops of aqua regia andevaporate to dryness on a water bath. Dissolve theresidue in water, neutralize, if required, with diluteammonium hydroxide and make up the volume to25 ml.

To’detect copper contamination, if any, in any of thereagents, blank experiment shall be carried out usingthe same quantities of the reagents.

15.3.3.1 Procedure (without extraction)

Take in 50-m\ Nessler tube, 10 mi of the test solutionprepared as described in 15.3.3. Add 2 g of citric acidand 10 ml of dilute ammonium hydroxide. Makeup to50 ml with water.

Prepare a series of control solutions, each containingin 50 ml, 2 g of citric acid and 10 ml of diluteammonium hydroxide together with an increasingamount of copper, namely, 0.1 ml, 0.2 ml, 0.4 ml,0.6 ml, 0.8 ml and 1.0 ml of standard copper solution(see 15.2.5).

The test solution and controls should be free from anyturbidity.

Cool all solutions to 20”C, add 2 ml ofdiethyldithiocarbamate solution to each and match thetest solution against the control solution. Note thenumber of millilitres of standard copper solution addedin the control of the test solution having, as nearly aspossible, the same intensity of colour as that of thetest solution.

15.3.3.2 Procedure (with extraction)

Extract immediately the copper organometalliccompound produced as described in the last paragraphunder 15.3.3.1 with four successive portions, 2.5 mleach, of carbon tetrachlorjde and compare the colourof the solution so obtained in a calorimeter with theextracts of control solution similarly prepared.

Chloroform may be used but carbon tetrachloride isbetter as it is almost insoluble in water and formsclearer solution, which separtifes quickly.

15.3.4 Calculation

Calculate copper as follows:

Copper (as Cu), in ppm = 0.2845 x 12.5 V

where

V= volume of standard comer solution in thereferee method is required in case of dispute between control solution which gives the closest match,the purchaser and the supplier. in ml.

8

.’,

,.

15.4 Potassium Ferrocyanide Method

15.4.1 Procedure

Transfer 20 ml of the material into silica evaporatingdish and add 1 ml of dilute sulphuric acid. Heat gentlyin the beginning and then evaporate almost to drynesson a water-bath. Ignite the residue over a smokelessflame to eliminate sulphuric acid. Cool, dissolve theresidue in 2 ml of water, add three drops of aqua regiaand evaporate to dryness on a water bath. Dissolvethe residue in 2 ml of dilute hydrochloric acid andwarm gently till the residue is dissolved. Add 0.5 g ofammonium chloride and dilute to 15 ml with waterdistilled in an all-glass apparatus. Add diluteammonium hydroxide till alkaline, Boil off excess ofammonia and filter into a clean Nessler tube. Cooland then render the solution acidic with acetic acid (3to 5 drops are usually sufficient). Dilute to 40 ml. Add0.5 ml of potassium ferrocyanide solution, stir andmake up the volume .to 50 ml.

NOTE — If the copper is more, a lesser amount, say 10 ml of

the material may be taken for the test.

15.4.1.1 Prepare a series of control solutions eachcontaining in 50 ml, 0.5 g of ammonium chloride, 3to 5 drops of acetic acid and 0.5 ml of potassiumferrocyanide solution together with an increasingamount of copper, namely, 2 ml, 4 ml, 6 ml, 8 ml and10 ml of the standard copper solution.

Compare the test solution (see 15.4.1) with controlsolutions and note the number ofmillilitres of standardcopper solution added in the control of the test solutionhaving, as nearly as possible, the same intensity ofcolour as that of the test solution.

15.4.2 Calculation

Calculate copper as follows:

Copper (as Cu), in ppm = 0.2845 x 12.5 V

where

V= volume of standard copper solution in thecontrol solution which gives the closest match,in ml.

16 DETERMINATION OF METHYL ALCOHOL

Two methods, namely, spectrophotometric method andgas chromatography method are employed. Thespectrophotometric method is suftlciently sensitive forroutine type of analysis. The gas chromatographymethod is more sensitive and shall serve as analternative method.

16.1 Spectrophotometric Method

16.1.1 Apparatus

16.1.1.1 Spectrophotometer, of any make with

IS 3752:2005

wavelength range from 350 to 700 nm and a maximumband width of 5 nm.

16.1.2 Reagents

16.1.2.1 Sodium bisulphite, AR grade.

16.1.2.2 Potassium permanganate solution —Dissolve 3.0 g of potassium permanganate and 15.0ml of phosphoric acid in 100 ml distilled water. Thesolution shall be prepared monthly.

16.1.2.3 Sodium salt of chromotropic acid solution(Sodium 1,8-dihydroxynaphthalene-3,6-disulphonateJ5 percent aqueous solution (m/v).

a)

b)

Prepare fresh solution every week of either acidor salt and filter, if not clear.

If necessary, prepare purified chromotropic acidby dissolving 10 g of chromotropic acid or itssodium salt in 25 ml water (add 2 ml sulphuricacid to aqueous solution of salt to convert it totlee acid). Add 50 ml of methyl alcohol heatjust to boiling and filter. Add 100 ml isopropylalcohol to precipitate free acid chromotropicacid. Add more isopropyl alcohol to increaseyield of purified acid.

16.1.2.4 Methanol stock solution — Dilute 1.0 g ofmethanol (99.9 percent, v/v) to 100 ml with 40 percent(v/v) ethanol (methanol free). Dilute 10 ml of thissolution to 100 ml with 40 percent ethanol (methanolfree).

16.1.2.5 Methanol standard solutions — Diluteappropriate volume of methanol stock solution(see 16.1.2.4) to 100-ml volumetric flasks with40 percent (v/v) ethanol (methanol free) to get finalconcentration 20,40,60,80 and 100 ppm of methanol.

16.1.3 Procedure

Take 50 ml of sample in a simple still and distil,collecting about 40 ml of distillate. Dilute 1 ml ofdistillate to 5 ml with distilled water and shake well.Take 1 ml of this solution, 1 ml of distilled water (forblank) and 1ml of each of methanol standard solutionsin to 50-ml stoppered test tubes and keep them in anice-cold water-bath. Add to each test tube 2 ml ofpotassium permangartate reagent and keep aside for30 min. Decolonize the solution by adding a littlesodium bisulphite and add 1 ml of chromotropic acidsolution. Mix well and add 15 ml of sulphuric acidslowly with swirling and place in hot water bathmaintaining at 80”C for 20 min. Observe the colourdevelopment from violet to red. Cool the reactionmixture and measure the absorbance at 575 nm using1 cm path length cell.

16.1.4 Calculation

Calculate methanol content in #l 00 Iitres of absoluteethanol as follows:

, t.,

‘1

9

LS 3752:2005

A2xcx~xloooxl(X)xloo”

Methanol =/4, .S

where

A,=

c.

D.

A, =

s.

absorbance for sample standard solution;

concentration of methanol standard solution,gfml;

dilution factor for sample solution;

absorbance for methanol standard solution; and

ethanol content of liquor sample in percent(v/v).

16.2 Gas Chromatographic Method

16.2.1 Apparatus

a) Gas chromatography and operating para-meters — Gas chromatography equipped withflame ionization detector and split injection portand fixed with a capillary column of HPCarbowax 20M or equivalent having thedimensions of 25 m length, 0.32 mm ID and0.30 y film thickness. The split ratio will beapproximately 1 : 40 with nitrogen or heliumas a carrier gas at the flow rate of about 1.7 ml/min. The detector and injector port temperaturesmay be maintained at about 250°C. Keep theoven temperature at 45°C for 4 rein, raise to10O°C/min at the rate of 10°C/min and finallyto 200°C for 10 min at the rate of 15°C

NOTE — Optimum operating conditions may vary with

co] umn and instrument used and must be determined by

using standard solutions. Adjust the parameters for maximum

peak sharpness and optimum separation. With high level

standard, rr-proprmol should give almost complete baseline

separation from ethanol.

b) Syringe — 10pl, Hamilton Co. No. 701, orequivalent.

16.2.2 Reagents

a) Ethanol — Methanol-free.

b) N-Pentanol internal standard — 0.05 percentWIV n-pentanol in 40 percent VIVethanol(methanol-free).

:.,.,,

itb, j

c) MethanoI stock solution — Dilute 1.0 g ofmethanol (99.9 percent, v/v) to 100 ml with40 percent (v/v) ethanol (methanol-free).

d) Methanol standard solution — Dilute 10 ml ofmethanol stock solution [see 16.2.2(c)] to100 ml with 40 percent (v/v) ethanol (methanol-tlee). Dilute 10 ml of this solution to 100 mlwith 40 percent (v/v) ethanol (methartol-free).Transfer 5 ml of the resulting solution intoa 10 ml-stoppered test tube, add 1 ml ofn-pentanol internal standard solution and mixwell.

16.2.3 Procedure

Transfer 5 ml of sample into a 10-ml stoppered testtube, add 1 ml of n-pentanol internal standard solutionand mix well. Inject 2pl of methanol itandard solutioninto chromatography and record the chromatogram.Adjust the operating parameters and attenuation toobtain measurable peaks (at least 25 percent of fill-scale deflection). Determine the retention time ofmethanol and n-pentanol. Inject 2pl sample solution [

.,”

into chromatography and record the chromatogram f:”

(adjust attenuation, if necessmy).~:,

16.2.4 Calculation

Calculate methanol content in grams per 100 litres ‘of $absolute alcohol as follows: ‘k-”

R2XCXDX l(J)OX1OOXI( )()‘.

Methanol =R,x S )

where

Rz = peak ratio of methanol to n-pentanol for sample )solution;

c.

D=

R,=

s=

concentration methanol in standard solution, ing/ml; .dilution factor for sample solution;

peak ratio of methanolstandard solution; and

ethanol content of liquor(v/v).

to n-pentanol for ‘4

sample, in percent

10

.

nl,IS 3752:2005

ANNEX AJ,,*;<$-

(Clause 1)i.,

ESTIMATION OF ESTERS, HIGHER ALCOHOLS, ALDEHYDES, FURFURAL AND

k

~:j.,,;

METHANOL BY GAS CHROMATOGRAPHIC METHOD{

A-1 DETAILED GAS CHROMATOGRAPHICMETHOD

A-1.l Apparatus

A-1.1.1 Gas Chromatography and OperatingParameters — Gas chromatography equipped withflame ionization detector and split injection port andfixed with a capillary column of HP Carbowax 20Mor equivalent having the dimensions of 25 ml length,0.32 mm ID and 0.30 p film thickness. The split ratiowill be approximately 1 :40 with nitrogen or heliumas a carrier gas at the flow rate of about 1.7 ml/min.The detector and injector port temperatures may bemaintained at about 250W. Keep the oven temperatureat 45°C for 4 rein, raise to 100W at the rate of 10°C/min and finally to 200”C for 10 min at the rate of 15°C/min.

NOTE—Optimum opmting conditions may vary with column

and instrument used and must be determined by using standard

solutions. Adjust the parameters for maximum peak sharpness

and optimum separation. With high level standard, rr-proparrol

should give almost complete baseline separation tlom ethanol.

A-1. 1.2 Syringe — 10 vI; Hamilton Co. No. 701, orequivalent.

A-1. 1.3 Reagentst

1) Internal standard: 0.5 percent (v/v) n-pentanolin 40 percent (v/v) ethanol (methanol-free).

I 2) Ethanol — Methanol-free.

} 3) Methanol

4) Acetaldehyde

5) IsobutyraldehydeI

I

6) Methyl acetate

7) Ethyl acetate

8) lso-valeraldehyde

9) n-Propyl acetate

1

10) Diacetyl

11) t-Amyl alcohol

12) n-Butyl acetate

13) Ethyl propionateL

14) n-Propanol

15) Iso-butanol

16) Iso-amyl acetate

17) n-Butanol

18) lso-amyl alcohol

19) Ethyl caprylate

I

20) Furfural

21) Ethyl caprate

22) Ethyl laurate

23) Phenethyl acetate

24) Phenethyl alcohol

25) Ethyl palmitate

26) Isovaleric acid

27) Ethyl caproate

28) Ethyl lactate

29) Acetic acid

30) Isobutyric acid

31) Phenyl acetate

32) Ethyl myristate

33) Caprylic acid

34) Pelargonic acid

35) Capric acid .,

A-1.1.4 Preparation of Standard Mixture *

:YTransfer accurately a known quantity of about 5.0 g ofthe reagents listed from A-1.1.3(3) to A-1.1.3(35) into different 100-ml volumetric flasks and dilute to i100 ml with 40 percent (v/v) ethanol (methanol-free).Transfer 1.0 ml of each of the resulting solutions intoa 100-ml volumetric flask and dilute to volume with40 percent (v/v) ethanol (methanol-free). This solution

)

will give approximately 500 ppm of each of componentlisted above.

.A-1. 1.4.1 Preparation of working standard mixture

Transfer 5 ml of standard mixture (see A-1.1.4) into a %10 ml-stoppered test tube, add 1ml of internal standardsolution [see A-1.1.3 (1)] and mix well.

A-1. 1.5 Procedure

Transfer 5 ml of sample into a 10 ml-stoppered testtube, add 1 ml of n-pentanol internal standard solutionand mix well. Inject 2@ of working standard mixture isolution into chromatography and record thechromatogram. Adjust the operating parameters andattenuation to obtain measurable peaks (at least 25percent of full-scale deflection). Determine theretention time of methanol and n-pentanol. Inject 2plsample solution into chromatography and record thechromatogram (adjust attenuation, if necessary).

NOTE — Identify the individual components by injecting

respective component standard solutions to the gas

chromatography and record the retention times.

11

IS 3752:2005

A-1. 1.6 Calculation

Calculate the individual component in grams per 100Iitres of absolute alcohol as follows:

Individual R2xCx~x1000x100xloo

component =R,x S

where

R2 = peak ratio of respective individual component(with respect to standard) to n-pentanol forsample solution;

C = concentration of respective individualcomponent in standard solution, in g/ml;

D = dilution factor for sample solution;

RI = peak ratio of respective individual componentto n-pentanol for standard solution; and

S = ethanol content of liquor sample in percent(v/v).

A-2 ROUTINE GAS CHROMATOGRAPHICMETHOD

A-2.1 Apparatus

A-2. 1.1 Gas Chromatography and Operating Para-meters — Gas chromatography equipped with flameionization detector and packed inlet and fixed with aglass column packed with 5 percent Carbowax 20Mon carbopak B, 80/120 mesh or equivalent packedcolumns like Porapak - Q having the dimensions of2 m in length and 4 mm in ID. Nitrogen or heliummay be used as carrier gas at suitable flow rate. Thedetector and injector port temperatures may bemaintained at about 2500C. Keep the oven temperatureat 45°C for 4 rein, raise to 100”C at the rate of 10°C/min and finally to 200”C for 10 min at the rate of 15°C/min.

NOTE — Optimum operating conditions may vary with column

and instrument used and must be determined by using standard

solutions. Adjust the parameters for maximum peak sharpness

and optimum separation. With high level standard, tr-proprmol

should give almost complete baseline separation from ethanol.

A-2.1.2 Syringe — Opl, Hamilton Co. No. 701, orequivalent.

A-2.1.3 Reagents

1)

2)3)4)5)6)7)8)9)

1o)

Internal standard: 0.5 percent (v/v) n-pentanolin 40 percent (v/v) ethanol (methanol-free).Ethanol — Methanol-free.MethanolAcetaldehydeEthyl acetaten-PropanolIso-butanolIso-amyl acetateIso-amyl alcoholEthyl caprylate

11) Furfural12) Ethyl caprate13) Ethyl Iaurate14) Phenethyl alcohol15) Ethyl caproate16) Ethyl lactate17) Acetic acid

A-2.1.4 Preparation of Standard Mixture

Transfer accurately a known quantity of about 5.0 gof the reagents listed from A-2.1.3(3) to A-2.1.3( I7)in to different 100 ml volumetric flasks and dilute to100-ml with 40 percent (v/v) ethanol (methanol-free).Transfer 1.0 ml of each of the resulting solutions intoa 100-ml volumetric flask and dilute to volume with40 percent (v/v) ethanol (methanol-free). This solutionwill give approximately 500 ppm of each of componentlisted above.

A-2.1.4.1 Preparation of working standard mixture

Transfer 5 ml of standard mixture (see A-2.1.4) ~to a10-ml stoppered test tube, add 1 ml of internal standardsolution [see A-2.1.3(1 )] and mix well.

A-2.1.5 Procedure

Transfer 5 ml of sample into a 10-ml stoppered testtube, add 1 ml of n-pentanol internal standard

ssolution and mix well. I~ject 2pl of working standardmixture solution into chromatography and record the 3’4

chromatogram. Adjust the operating parameters andattenuation to obtain measurable peaks (at least 25percent of full-scale deflection). Determine the i

retention time of methanol and n-pentanol. Inject 21.dsample solution into chromatography and record thechromatogram (adjust attenuation, if necessary). >

NOTE — Identify the individual components by injecting

respective component standard solutions to the gas

chrornatograph and record the retention times.

A-2.1.6 Calculation

Calculate the individual component in grams per 100 ‘4litres of absolute alcohol as follows:

Individual ~,xcx~xloooxlooxloo

component =R,x S

where

Rz = peak ratio of respective individual component(with respect to standard) to n-pentanol for

i

sample solution;

C = concentration of respective individual.

component in standard solution in g/ml;

D = dilution factor for sample solution;

R,= peak ratio of respective individual componentto n-pentanol for standard solution; and

S = etahnol content of liquor sample in percent(v/v).

12

IS 3752:2005.’,

[S No.

264:1976

265:1987

266:1976

1070:1992

ANNEX B

(C)au.se 2)

LIST OF REFERRED INDIAN STANDARDS

Title

Nitric acid — Specification(second revision)

Hydrochloric acid —Specification ~ourth revision)

Specification ‘for sulphuric acid(third rmision)

Reagent grade water —Specification (third revision)

IS No. Title

2263:1979 Methods of preparation ofindicator solutions (firstrevision)

2302:1989 Tables for alcoholometry @strevision)

3506:1989 Tables for alcoholometry (byPyknometer method) (firstrevision)

13

,

Bureau of Indian Standards

BIS is a statutory institution established under the Bureau of Indian Standards Act, 1986 to promoteharmonious development of the activities of standardization, marking and quality certification of goods andattending to connected matters in the country.

Copyright

BIS has the copyright of all its publications. No part of these publications may be reproduced in any formwithout the prior permission in writing of BIS. This does not preclude the free use, in the course of implementingthe standard, of necessary details, such as symbols and sizes, type or grade designations. Enquiries relating tocopyright be addressed to the Director (Publications), BIS.

Review of Indian Standards

Amendments are issued to standards as the need arises on the basis of comments. Standards are also reviewedperiodically; a standard along with amendments is reaffirmed when such review indicates that no changes areneeded; if the review indicates that changes are needed, it is taken up for revision. Users of Indian Standardsshould ascertain that they are in possession of the latest amendments or edition by referring to the latest issue of‘BIS Catalogue’ and ‘Standards: Monthly Additions’.

This Indian Standard has been developed from Dot: No. FAD 14 (1532).

Amendments Issued Since Publication

Amend No. Date of Issue Text Affected

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